The present invention is directed to a method of preparing an artificial tooth primordium in vitro, comprising the steps: a) providing isolated mesenchymal dental pulp cells; and b) culturing the mesenchymal dental pulp cells under non-adherent conditions to form a cell aggregate representing an artificial tooth primordium; as well as to an artificial tooth primordium derived therefrom.

A dental prosthesis to be integrated into a jaw bone cavity of a pre-identified patient. An example of a dental prosthesis includes a first manufactured portion having a surface shaped to substantially dimensionally conform three dimensionally to an undersized shape of the outer three-dimensional surface shape of a root of a tooth to be replaced, and a second manufactured portion shaped to substantially conform to the three-dimensional surface of a crown of the tooth. The outer surface of the root portion can include or be coated with a biocompatible material that is suitable to be integrated into the extraction socket and adopted by existing tissue forming the socket.

Provided is a dental pretreatment material for dental tissue regeneration by use of dental pulp stem cells, particularly a dental pretreatment material effectively enabling dental tissue regeneration even by use of dental pulp stem cells of middle-aged or older individuals. The dental pretreatment material is characterized by comprising a serine protease, specifically trypsin. The dental pretreatment material comprising trypsin is used as an injection into a root canal before a root canal filling material comprising dental pulp stem cells and an extracellular matrix is inserted into the root canal as an attempt to regenerate a dental pulp and a dentin. The root canal filling material includes an ALK5 inhibitor, a CCR3 antagonist, or a CCL11 neutralizing antibody.

Methods of manufacturing dental prosthesis/implants each to replace a non-functional natural tooth positioned in a jawbone of a specific pre-identified patient are provided. An example method includes the steps of receiving imaging data such as x-ray image data and surface scan data of a dental anatomy and/or a physical impression of the dental anatomy of a specific preidentified patient. The steps can also include forming a three-dimensional virtual model of at least portions of a non-functional natural tooth positioned in the jawbone of the specific pre-identified patient based on the imaging and surface scan data, virtually designing a dental implant based upon the virtual model, exporting the data describing the designed dental implant to a manufacturing machine, and custom manufacturing the dental implant for the specific patient.

Methods of manufacturing dental prosthesis/implants each to replace a non-functional natural tooth positioned in a jawbone of a specific pre-identified patient are provided. An example method includes the steps of receiving imaging data such as x-ray image data and surface scan data of a dental anatomy and/or a physical impression of the dental anatomy of a specific preidentified patient. The steps can also include forming a three-dimensional virtual model of at least portions of a non-functional natural tooth positioned in the jawbone of the specific pre-identified patient based on the imaging and surface scan data, virtually designing a dental implant based upon the virtual model, exporting the data describing the designed dental implant to a manufacturing machine, and custom manufacturing the dental implant for the specific patient.

Disclosed is a root canal filler for non-extracted tooth which causes no internal resorption or external resorption in a tooth with complete root formation, shows no odontoclast, and contributes to the regeneration of a dental tissue in which odontoblasts are smoothly aligned on the dentin wall. After pulpectomy or enlargement/cleaning of an infected root canal, a root canal filler for non-extracted tooth, which comprises tooth pulp stem cells and an extracellular matrix, is inserted into the apical side of the root canal of the non-extracted tooth. The tooth pulp stem cells may be, for example, dental pulp CXCR4-positive cells. It is preferred to attach, to the crown side of the root canal, migration factor(s) including at least one factor selected from among a cell migration factor, a cell proliferation factor, a neurotrophic factor and an angiogenic factor.

Provided is a method for making a biotransplant, comprising introducing autologous fibroblasts isolated from an oral mucosa of a patient into a platelet-rich fibrin (PRF) membrane using linear retrograde needle injection, where a needle is inserted into a thickness of the PRF membrane and a first puncture is made and then a series of punctures are made that are linearly aligned or arrayed with the first puncture and are spaced at a predetermined distance from a prior puncture. A method of treatment of a periodontal tissue and a biotransplant comprising a PRF membrane and autologous fibroblasts are also provided.

The present disclosure provides a hydrogel composition comprising an interpenetrating polymer network (IPN) containing a biopolymer, a first synthetic polymer and a second synthetic polymer in which a contained community of live human MSCs is embedded. The collagen polymer matrix described (a) allows the embedded cells to remain in place or to migrate over short distances; (b) allows diffusion of small molecules, particularly growth factors produced by the cells or provided as a supplement, and EVs released by the cells to support the recovery of periodontium tissue function following injury.

Embodiments of the disclosure include means of treating a variety of periodontal and dental diseases using fibroblasts, modified fibroblasts, and derivatives thereof. In one embodiment, the disclosure encompasses the utilization of fibroblasts for treatment of gum disease. In another embodiment, disclosed are means of utilizing fibroblasts and modifications thereof, for preparing gum tissue for placement of an artificial tooth. In another embodiment, fibroblasts are utilized to increase efficacy of bone grafts. In other embodiments, fibroblasts are utilized to generate artificial teeth through utilization of various scaffolding and/or 3 dimensional printing means.

The present invention relates to a method of preparing a three-dimensional (3D) cell composition comprising the steps of a) forming a support matrix containing oral fibroblasts suspended within the support matrix by mixing fibrinogen, a modifier and oral fibroblasts with thrombin, b) incubating the support matrix in a cell culture media for a sufficient time to allow development of a first layer of the three-dimensional cell composition, and c) seeding oral keratinocytes on a surface of the first layer and culturing the oral keratinocytes to form a second layer of the three-dimensional cell composition. In specific embodiments of the invention, the method is exemplified for the production of an artificial gingival tissue, wherein polyethylene oxide), 4-arm, succinimidyl glutarate terminated is used as the modifier. Also disclosed are uses of the 3D cell composition which include treatment of a gum disease or condition, regenerative therapy and for in vitro testing.